The regulatory role of natural progesterone in barrier immunity

摘要：孕激素（P4）是调节月经生理变化的关键性激素 周期我们和其他人已经表明，它也有抗炎作用的免疫系统局部在 女性生殖道（FRT），但机制还不清楚。也有迹象表明， 其免疫作用超过了FRT，并影响免疫系统。P4的影响可能会被修改， 雌二醇（E2）的促炎作用更强。详细了解背后的机制 P4在局部和全身部位的免疫活性及其与E2的相互作用可能有助于 通过解释基于性别和月经周期相关的炎症性疾病的波动， 疾病一些炎症性疾病在月经周期中波动，这表明P4具有 全身免疫效应。例如，哮喘和炎症性肠病在P4-低水平时恶化， 卵泡期，而类风湿性关节炎往往在P4主导的黄体期改善。另一方面，在一项研究中， P4诱导的免疫抑制可能限制对疫苗接种的反应或对感染的抵抗。 P4在月经周期的黄体期激增，达到卵泡期水平的40倍以上。我们 数据显示在各阶段之间FRT的明显免疫学差异：巨噬细胞的黄体抑制- 趋化因子P4。单核/巨噬细胞和树突状细胞（DC）是抗原呈递细胞 (APC)，其为驻留记忆T细胞（TRM）的定位和存活提供关键信号。 我们提出了互补的人类和小鼠研究：（1）在APC水平进行研究的人类队列， 在FRT和其他屏障部位探索它们与TRM的联系，以及（2）小鼠实验以证实 P4与E2结合的免疫学特性，并检验阴道CCL 2/CCL 4 给药部分逆转P4诱导的免疫抑制。 在目标1中，我们将招募一个临床队列来研究人类P4主导黄体期的影响 对APC和TRM在FRT、胃肠道、上呼吸道、 皮肤和血液。我们将使用基于MSD的分泌物免疫因子分析和流式细胞术， 粘膜细胞的单细胞RNA测序以解决三个假设：（1）APC-嗜性的抑制 趋化因子的产生是黄体期屏障部位免疫抑制的标志;（2）粘膜免疫抑制 APC在黄体期比卵泡期具有更少或更少的炎性;以及（3）APC和细胞因子在黄体期比卵泡期具有更少或更少的炎性。 黄体期的变化与TRM的数量和/或激活状态降低有关。 在目标2中，我们将使用小鼠模型来建立P4治疗、趋化因子 生产，APC和TRM频率和激活状态。我们将讨论两个假设：（1）P4 抑制主要屏障组织中CCL 2和CCL 4的产生，降低APC和TRM频率，和/或 功能潜力;和（2）阴道CCL 2/4给药防止P4诱导的APC和TRM抑制。

ABSTRACT: Progesterone (P4) is a key sex hormone governing the physiological changes of the menstrual cycle. We and others have shown that it also has anti-inflammatory effects on the immune system locally in the female reproductive tract (FRT), but the mechanisms are not well understood. There are also indications that its immune effects reach beyond the FRT and affect immunity systemically. P4’s effects are likely modified by the more pro-inflammatory effects of estradiol (E2). A detailed understanding of the mechanisms behind P4’s immune activities at local and systemic sites, and its interactions with E2, may contribute to women’s health by explaining sex-based and menstrual cycle-related fluctuations in inflammatory diseases. Some inflammatory diseases fluctuate during the menstrual cycle, suggesting that P4 has systemic immune effects. For example, asthma and inflammatory bowel disease worsen in the P4-low follicular phase, while rheumatoid arthritis often improves in the P4-dominant luteal phase. On the other hand, P4-induced immunosuppression could limit the response to vaccination or resistance to infection. P4 surges during the luteal phase of the menstrual cycle, reaching >40 times above follicular phase levels. Our data show a clear immunological difference in the FRT between the phases: luteal inhibition of macrophage- tropic chemokines by P4. Monocytes/macrophages and dendritic cells (DC) are antigen-presenting cells (APC), which provide key signals for the localization and survival of resident memory T cells (TRM). We propose complementary human and mouse studies: (1) a human cohort with studies at the APC level, probing their connections to TRM in the FRT and other barrier sites and (2) mouse experiments to corroborate P4’s immunological properties in combination with E2 and to test the hypothesis that vaginal CCL2/CCL4 administration partially reverses P4-induced immune suppression. In Aim 1, we will recruit a clinical cohort to study the effects of the P4-dominant luteal phase in humans on APC and TRM activation and function in the FRT, the gastrointestinal tract, the upper respiratory tract, the skin, and the blood. We will use MSD-based immune factor profiling of secretions and flow cytometry and single cell RNA sequencing of mucosal cells to address three hypotheses: (1) inhibition of APC-tropic chemokine production is a hallmark of barrier site immunosuppression during the luteal phase; (2) mucosal APCs are fewer or less inflammatory during the luteal than the follicular phase; and (3) the APC and cytokine changes during the luteal phase associate with lower number and/or activation status of TRM. In Aim 2, we will use mouse models to establish a causal link between P4 treatment, chemokine production, and APC & TRM frequencies and activation state. We will address two hypotheses: (1) P4 inhibits production of CCL2 and CCL4 in major barrier tissues, reducing APC and TRM frequencies and/or functional potential; and (2) vaginal CCL2/4 administration prevents P4-induced suppression of APC and TRM.